May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Two-photon Excited Autofluorescence Imaging of the Human Retina-Choroid Complex
Author Affiliations & Notes
  • M. Han
    Mannheim Biomedical Engineering Labs., University of Heidelberg, Germany
  • G. Giese
    Max Planck Institute for Medical Research, Heidelberg, Germany
  • S. Schmitz-Valckenberg
    Department of Ophthalmology, University of Bonn, Germany
  • A. Bindewald-Wittich
    Department of Ophthalmology, University of Bonn, Germany
  • F. G. Holz
    Department of Ophthalmology, University of Bonn, Germany
  • M. H. Niemz
    Mannheim Biomedical Engineering Labs., University of Heidelberg, Germany
  • J. F. Bille
    Kirchhoff Institute for Physics, University of Heidelberg, Germany
  • Footnotes
    Commercial Relationships M. Han, None; G. Giese, None; S. Schmitz-Valckenberg, None; A. Bindewald-Wittich, None; F.G. Holz, None; M.H. Niemz, None; J.F. Bille, None.
  • Footnotes
    Support None.
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 4245. doi:
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      M. Han, G. Giese, S. Schmitz-Valckenberg, A. Bindewald-Wittich, F. G. Holz, M. H. Niemz, J. F. Bille; Two-photon Excited Autofluorescence Imaging of the Human Retina-Choroid Complex. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4245.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose:
 

The development of age-related macular degeneration (AMD) is thought to result from an accentuation of the aging process. A minimally invasive, high resolution imaging modality is vital to delineate the age-related structural abnormalities in the retina-choroid complex prior to apparent pathological manifestations of AMD. Based on the endogenous chromophores in the human retina-choroid complex, we employed two-photon excited autofuorescence (TPEF) imaging to characterize the morphological and spectral properties of the human photoreceptors, RPE and choriocapillaries ex vivo.

 
Methods:
 

The human retinas were obtained from four human Caucasian postmortem donor eyes (19, 55, 57 and 64 years old, with normal vision). TPEF imaging was performed with a customized multiphoton laser scanning microscope (Zeiss LSM 510) equipped with a femtosecond Ti:Sapphire laser (720-980 nm). The autofluorescence spectrum of RPE cells was measured with a confocal laser scanning microscope (Leica TCS SP2) coupled to a UV argon laser (364 nm).

 
Results:
 

As age increases, the regularity of the parafoveal photoreceptor mosaic appears to be preserved (Fig.a,b). Enlarged lipofuscin granules in the aged RPE demonstrate significantly blue-shifted autofluorescence (Fig.d), which coincides with the depletion of melanin pigments. Prominent fibrillar structures in elderly choriocapillaries may represent age-related structure and permeability alterations in the choroid (Fig.f).

 
Conclusions:
 

TPEF imaging is an elegant and highly efficient tool to delineate the thick, fragile and opaque retina-choroid complex. Requiring neither slicing nor labeling, the most vital components (photoreceptors, RPE, choriocapillaries, Bruch's membrane) in the human retina-choroid complex can be examined with high contrast and subcellular resolution, which may provide fresh insights into the pathogenesis of AMD.  

 
Keywords: aging • retina • choroid 
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